Endoplasmic reticulum molecular chaperone involved in the impairment of inner ear consistent with the mimetic aging rats / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the involvement of endoplasmic reticulum molecular chaperone GRP78 in the impairment of inner ear consistent with the mimetic aging model.@*METHOD@#Twenty-four Wistar rats were randomly divided into two groups. Model group was induced by daily hypodermic injection of 10% D-galactose (800 mg x kg(-1) x d(-1)) for 8 weeks and the control group was given saline accordingly. Spatial learning and memory was measured by Morris-Water-Maze. Colorimetry was used to analyze superoxide dismutase (SOD) and malondialdehyde (MDA) extracted from inner ear tissue. Hearing threshold of rats were detected with Auditory brainstem response (ABR). In addition, expression of GRP78 in the inner ear was detected by immunohistochemistry, RT-PCR and Western blot. The control group was studied parallel.@*RESULT@#The escape latency in the model group injected with D-galactose was markedly longer than that in the control group. Accordingly, the changes of SOD and MDA were more significant in the model group, the difference between two groups was significant (t-test, P0.05). The expression of GRP78 was significantly different between two groups, which is increased in the inner ear tissue of model group (P<0.01).@*CONCLUSION@#The impairment of inner ear tissue partly dued to the oxidative stress in the model, which was induced by D-galactose and endoplasmic reticulum molecular chaperone was thought to contribute to the impairment mechanism of inner ear in mimetic aging model.

Endoplasmic reticulum molecular chaperone involved in the impairment of inner ear consistented with the mimetic aging rats / 临床耳鼻咽喉头颈外科杂志

Objective:To explore the involvement of endoplasmic reticulum molecular chaperone GRP78 in the impairment of inner ear consistented with the mimetic aging model.Method:Twenty-four Wistar rats were randomly divided into two groups. model group was in duced by daily hypodermic injection of 10% D-galactose (800 mg·kg~(-1)·d~(-1)) for 8 weeks and the control group was given saline accordingly. Spatial learning and memory was measured by Morris-Water-Maze. Colorimetry was used to analyze superoxide dismutase (SOD) and malondialdehyde (MDA) extracted from inner ear tissue. Hearing threshold of rats were detected with Auditory brainstem response (ABR).In addition, expression of GRP78 in the inner ear was detected by immunohistochemistry,RT-PCR and Western blot. The control group was studied parallel.Result:The escape latency in the model group injected with D-galactose was markedly longer than that in the control group.accordingly ,the changes of SOD and MDA were more significant in the model group, the difference between two groups was significant(t-test,P0.05). The expression of GRP78 was significantly different between two groups ,which is increased in the inner ear tissue of model group(P<0.01).Conclusion:The impairment of inner ear tissue partly dued to the oxidative stress in the model, which was induced by D-galactose.and endoplasmic reticulum molecular chaperone was thought to contribute to the impairment mechanism of inner ear in mimetic aging model.

Study of the effect of JNK signal transduction pathway in intense noise-induced apoptosis in cochlea of guinea pig / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the mechanism of intense noise-induced cochlea cells death in guinea pig, and the effect of JNK signal transduction pathway in the procedure of cochlea cells apoptosis by intense noise-induced.@*METHOD@#Thirty-two guinea pigs were randomly divided into 4 groups. The guinea pigs in the experiment groups were exposed to 4 kHz narrow band noise at 120 dB SPL for 4 h. After the noise expose for 1, 4, 14 days of the experiment guinea pigs, ABR of the guinea pigs on experiment and control groups were tested before put them to death. Four guinea pig's cochleas of every group were taken to paraffin section, and the rest was extracted the total cochlear's protein. Apoptosis was tested by terminal deoxynucleotidyl Transferase (TdT)-mediated deoxyuridine triphosphate (d-UTP) nick and labeling method (TUNEL). The phosphorylation of JNK and c-Jun were tested by immunohistochemistry and western blot methods.@*RESULT@#Tunel-Positive cells in the Corti's, SGC and SV of experiment groups, and there have significant differences compared with the control group (P<0.01) and Tunel-Positive cells are most in 1 d experiment group. The positive cells of P-JNK and P-c-Jun could be detected in guinea pig's cochleas after noise exposed, but no positive cells were found in the control. Protein levels of P-JNK and P-c-Jun were risen up and activated quickly after noise exposed, and achieved peak in 1 d, 4 d and then fallen-offs, but still maintained higher levels within 14 d.@*CONCLUSION@#Intense noise causes cochlea cell lesion by inducing apoptosis to result in and JNK signal transduction pathway plays an important role in the procedure of apoptosis.

Study of the effect of JNK signal transduction pathway in intense noise-induced apoptosis in cochlea of guinea pig / 临床耳鼻咽喉头颈外科杂志

Objective:To investigate the mechanism of intense noise-induced cochlea cells death in guinea pig,and the effect of JNK signal transduction pathway in the procedure of cochlea cells apoptosis by intense noise-induced.Method:Thirty-two guinea pigs were randomly divided into 4 groups.The guinea pigs in the experiment groups were exposed to 4 kHz narrow band noise at 120 dB SPL for 4 h.After the noise expose for 1,4,14 days of the experiment guinea pigs,ABR of the guinea pigs on experiment and control groups were tested before put them to death.Four guinea pig's cochleas of every group were taken to paraffin section,and the rest was extracted the total cochlear's protein.Apoptosis was tested by terminal deoxynucleotidyl Transferase(TdT)-mediated deoxyuridine triphosphate(d-UTP)nick and labeling method(TUNEL).The phosphorylation of JNK and c-Jun were tested by immunohistochemistry and western blot methods.Result:Tunel-Positve cells in the Corti's,SGC and SV of experiment groups,and there have significant differences compared with the control group(P＜0.01)and Tunel-Positve cells are most in 1 d experiment group.The positive cells of P-JNK and p-e-Jun could be dectected in guinea pig's cochleas after noise exposed,but no positive cells were found in the control.Protein levels of p-JNK,and P-c-Jun were risen up and activated quickly after noise exposed,and achieved peak in 1 d,4 d,and then fallen-offs,but still maintained higher levels within 14 d.Conclusion:Intense noise causes cochlea cell lesion by inducing apoptosis to result in and JNK signal transduction pathway plays an important role in the procedure of apoptosis.

Effect of the intense noise on hearing function and cochlea morphology in rat / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the changes of auditory electrophysiology and inner ear pathology in rat cochlea after the noise exposure, and to offer the experimental data for exploring the mechanism of noise-damaged cochlea.@*METHOD@#The rats in the study group were exposed to a intense narrow band noise centered at 4 kHz at the leave of 120 dB (SPL) for 4 h. The exposed cochleae were collected at various intervals (1 or 21 days) after the noise exposure. Auditory function was monitored by measuring thresholds of auditory brain stem responses (ABR). The morphological changes in rat cochlear hair cell (HC) were examined by HC nuclei stained with Propidium iodide (PI), a fluorescent dye specifically labelling the nuclear DNA and scanning electron microscopy (SEM). The number of spiral ganglion cells was calculated using pathologic technique.@*RESULT@#The thresholds of ABR in the study group were significantly greater than that in the normal control group (P 0.05). SEM revealed the injured stereocilia of OHC (disarrangement, collapse) and OHC loss in the study group, which was more severe in OHC3 than the other two rows of OHC.@*CONCLUSION@#The intense noise used in our study could injure the rat cochlea and bring permanent threshold shift (PTS). Under this condition, the death modes of HC in the cochlea include apoptosis and necrosis in the fore part, whereas necrotic is the major mode in the evening of exposure. The injured stereocilia of OHC and OHC loss could remain the most consistent correlate of PTS.

Effect of DRB on the Biological Characteristics of Human Laryngeal Carcinoma Hep-2 Cell Line / 华中科技大学学报（医学）（英德文版）

In order to study the effect of 5, 6-Dichloro-1-β-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68±0.19) %, (1.95±0.12)%, (8.51 ±0.26)%, (11.26±0.17) % and (14.99±0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.

Culture, identification and label of embryonic rat neural stem cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the methods of culture, identification and label of embryonic rat neural stem cells.@*METHOD@#The cells isolated from fetal rat hippocampus were identified with nestin immunocytochemical fluorescent staining. The cellular multiplication was observed by immunocytochemical fluorescence co-label after accession of BrDU. The neural stem cells (NSCs) were marked with fluorescent dye, bisbenzimide (Hoechest33342) and induced to differentiate. The differentiated cells were detected with Neuron Specific Enolase (NSE) and Glial Fibrillary Acidic Protein (GFAP) immunocytochemical fluorescent staining respectively.@*RESULT@#Nest-like clusters of neural stem cells were obtained in suspension and the cells could be differentiated into neurons and astrocytes which maintaining the main characteristics of NSCs after 8 passages of culture. The label efficiency of cells with Hoechest33342 was 97% and no attenuation of fluorescent brightness was observed after 8 passages of culture. The cellular fluorescence was observed in the NSCs and the differentiated cells.@*CONCLUSION@#The cells from embryonic rat hippocampus possessed the abilities of division, multiplication and self-renew, which were believed to be the main characteristics of NSCs of the central nervous system. The cells could be efficiently labeled with fluorescent dye and could be used as donor cells in experimental research on NSCs transplantation.

Effect of DRB on the biological characteristics of human laryngeal carcinoma Hep-2 cell line / 华中科技大学学报（医学）（英德文版）

In order to study the effect of 5, 6-Dichloro-1-beta-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose-and time-dependent manner. After being treated with 0, 10, 20, 40, 80 microm mol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68+/-0.19)%, (1.95+/-0.12)%, (8.51+/-0.26)%, (11.26+/-0.17)% and (14.99+/-0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 microm mol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.

The study of TNF-α destructive action to the bone in human middle cholesteatoma / 临床耳鼻咽喉头颈外科杂志

Objective:To study the expression of TNF-α in the cholesteatoma and elucidate the role of TNF-α in the destruction of bone.Method:All samples (5 μm paraffin sections) from 25 cholesteatoma cases and 10 normal cases were examined by immunohistochemical SABC method and analysed by computer image.Result:In 25 cholesteatoma cases,TNF-α expressed in the cytoplasm of the full epithelial tissue cells,subcutaneous inflammatory cells and fibroblast.In contrast,the normal cases,there were 6 cases week positive,4 cases negative.The results of the computer image quantitative analysis system showed that the mean optical density of TNF-α was 0.1326±0.0022 in the cholesteatoma epithelial tissue and 0.0868±0.0014 in normal skin epithelial tissue respectively,both of which had significantly differentiation (P＜0.05).Conclusion:TNF-α may be a factor of the destruction of bone of cholesteatoma.

Clinical Study on Treatment of 56 Cases of Sequelae of Cerebral Hemorrhage with Fu Fang Qi Dan Dai Zhu San / 中医杂志

Objective:To observe clinical therapeutic effect of Fu Fang Qi Dan Dai Zhu San on sequelae of cerebral hemorrhage. Methods: 56 cases were selected randomly from the 315 cases who had received functional exercise, massage and other rehabilitation treatments. The patient were administrated by Fu Fang Qi Dan Dai Zhu San Decoction for 3 months, which is constituted by Bu Yang Huan Wu Decoction and Fu Fang Dan Shen Tablets, and the therapeutic effect was compared with that of Bu Yang Huan Wu Decoction, Fu Fang Dan Shen Tablets and other clinically commomly - used drugs, respectively. Results: Both the clinically cured rate and the total markedly effective rate in the treatment group of Fu Fang Qi Dan Dai Zhu San were significantly higher than those of the six control groups. Conclusion: Fu Fang Qi Dan Dain Zhu San can obviously increase the therapeutic effect and shorten the therapeutic course, and it is a good prescription for treatment of sequelae of cerebral hemorrhage.